空间站微重力流体实验设备需求分析

对国际空间站和中国科学实验卫星及载人飞行器上开展的微重力流体实验情况进行论述和分析,重点分析了国际空间站（ISS）微重力流体科学实验设备情况.根据中国空间微重力流体物理科学发展需求,结合国际空间站微重力流体科学实验对设备的需求,提出了未来在中国空间站开展微重力流体实验时空间实验设备需要重点考虑和解决的问题,同时提出相关设计建议.      

Seed-to-seed growth of Arabidopsis thaliana on the International Space Station.

The assembly of the International Space Station (ISS) as a permanent experimental outpost has provided the opportunity for quality plant research in space. To take advantage of this orbital laboratory, engineers and scientists at the Wisconsin Center for Space Automation and Robotics (WCSAR), University of Wisconsin-Madison, developed a plant growth facility capable of supporting plant growth in the microgravity environment. Utilizing this Advanced Astroculture (ADVASC) plant growth facility, an experiment was conducted with the objective to grow Arabidopsis thaliana plants from seed-to-seed on the ISS. Dry Arabidopsis seeds were anchored in the root tray of the ADVASC growth chamber. These seeds were successfully germinated from May 10 until the end of June 2001. Arabidopsis plants grew and completed a full life cycle in microgravity. This experiment demonstrated that ADVASC is capable of providing environment conditions suitable for plant growth and development in microgravity. The normal progression through the life cycle, as well as the postflight morphometric analyses, demonstrate that Arabidopsis thaliana does not require the presence of gravity for growth and development.     

Improvements in the re-flight of spaceflight experiments on plant tropisms

In order to effectively study phototropism, the directed growth in response to light, we performed a series of experiments in microgravity to better understand light response without the “complications” of a 1-g stimulus. These experiments were named TROPI (for tropisms) and were performed on the European Modular Cultivation System (EMCS), a laboratory facility on the International Space Station (ISS). TROPI-1 was performed in 2006, and while it was a successful experiment, there were a number of technical difficulties. We had the opportunity to perform TROPI-2 in 2010 and were able to optimize experimental conditions as well as to extend the studies of phototropism to fractional gravity created by the EMCS centrifuge. This paper focuses on how the technical improvements in TROPI-2 allowed for a better experiment with increased scientific return. Major modifications in TROPI-2 compared to TROPI-1 included the use of spaceflight hardware that was off-gassed for a longer period and reduced seed storage (less than 2 months) in hardware. These changes resulted in increased seed germination and more vigorous growth of seedlings. While phototropism in response to red illumination was observed in hypocotyls of seedlings grown in microgravity during TROPI-1, there was a greater magnitude of red-light-based phototropic curvature in TROPI-2. Direct downlinking of digital images from the ISS in TROPI-2, rather than the use of analog tapes in TROPI-1, resulted in better quality images and simplified data analyses. In TROPI-2, improved cryo-procedures and the use of the GLACIER freezer during transport of samples back to Earth maintained the low temperature necessary to obtain good-quality RNA required for use in gene profiling studies.     

Simulations of absorbed dose on the phantom surface of MATROSHKA-R experiment at the ISS

The health risks associated with exposure to various components of space radiation are of great concern when planning manned long-term interplanetary missions, such as future missions to Mars. Since it is not possible to measure the radiation environment inside of human organs in deep space, simulations based on radiation transport/interaction codes coupled to phantoms of tissue equivalent materials are used. However, the calculated results depend on the models used in the codes, and it is therefore necessary to verify their validity by comparison with measured data. The goal of this paper is to compare absorbed doses obtained in the MATROSHKA-R experiment performed at the International Space Station (ISS) with simulations performed with the three-dimensional Monte Carlo Particle and Heavy-Ion Transport code System (PHITS). The absorbed dose was measured using passive detectors (packages of thermoluminescent and plastic nuclear track detectors) placed on the surface of the spherical tissue equivalent phantom MATROSHKA-R, which was exposed aboard the ISS in the Service Zvezda Module from December 2005 to September 2006. The data calculated by PHITS assuming an ISS shielding of 3 g/cm² and 5 g/cm² aluminum mass thickness were in good agreement with the measurements. Using a simplified geometrical model of the ISS, the influence of variations in altitude and wall mass thickness of the ISS on the calculated absorbed dose was estimated. The uncertainties of the calculated data are also discussed; the relative expanded uncertainty of absorbed dose in phantom was estimated to be 44% at a 95% confidence level.     

Inhibitory effect of simulated microgravity on differentiating preosteoblasts

The bone loss induced by microgravity is partly due to the decrease of mature osteoblasts. In the present study, we employed the random positioning machine (RPM) to simulate microgravity and investigated the acute effects of simulated microgravity on the differentiation of 2T3 preosteoblasts. Following 7 days’ culture under normal (1 g) condition, cells were exposed to simulated microgravity for 24 h. The results showed that 24 h treatment of simulated microgravity significantly decreased alkaline phosphatase (ALP) activity without changing the cell morphology. In addition, the mRNA expressions of osteogenic genes, including runt-related gene 2 (Runx2), osterix, osteocalcin (OC), type I collagen (Col I) and bone morphogenetic protein (BMP), were dramatically downregulated. Moreover, western blot analysis of total extracellular signal-regulated kinase (Erk) and phosphorylated Erk (p-Erk) indicated that p-Erk level, which represents the Erk activation status, was increased. Taken together, our results suggested that acute exposure to simulated microgravity inhibited osteoblast differentiation through modulating the expression of osteogenic genes and the Erk activity. These findings provide new insight for bone loss due to microgravity and unloading.     

"From seed-to-seed" experiment with wheat plants under space-flight conditions.

An important goal with plant experiments in microgravity is to achieve a complete life cycle, the "seed-to-seed experiment." Some Soviet attempts to reach this goal are described, notably an experiment with the tiny mustard, Arabidopsis thaliana, in the Phyton 3 device on Salyut 7. Normal seeds were produced although yields were reduced and development was delayed. Several other experiments have shown abnormalities in plants grown in space. In recent work, plants of wheat (Triticum aestivum) were studied on the ground and then in a preliminary experiment in space. Biometric indices of vegetative space plants were 2 to 2.5 times lower than those of controls, levels of chlorophyll a and b were reduced (no change in the ratio of the two pigments), carotenoids were reduced, there was a serious imbalance in major minerals, and membrane lipids were reduced (no obvious change in lipid patterns). Following the preliminary studies, an attempt was made with the Svetoblock-M growth unit to grow a super-dwarf wheat cultivar through a life cycle. The experiment lasted 167 d on Mir. Growth halted from about day 40 to day 100, when new shoots appeared. Three heads had appeared in the boot (surrounded by leaves) when plants were returned to earth. One head was sterile, but 28 seeds matured on earth, and most of these have since produced normal plants and seeds. In principle, a seed-to-seed experiment with wheat should be successful in microgravity.     

Improvements in and actual performance of the Plant Experiment Unit onboard Kibo,the Japanese experiment module on the international space station

In 2004, Japan Aerospace Exploration Agency developed the engineered model of the Plant Experiment Unit and the Cell Biology Experiment Facility. The Plant Experiment Unit was designed to be installed in the Cell Biology Experiment Facility and to support the seed-to-seed life cycle experiment of Arabidopsis plants in space in the project named Space Seed. Ground-based experiments to test the Plant Experiment Unit showed that the unit needed further improvement of a system to control the water content of a seedbed using an infrared moisture analyzer and that it was difficult to keep the relative humidity inside the Plant Experiment Unit between 70 and 80% because the Cell Biology Experiment Facility had neither a ventilation system nor a dehumidifying system. Therefore, excess moisture inside the Cell Biology Experiment Facility was removed with desiccant bags containing calcium chloride. Eight flight models of the Plant Experiment Unit in which dry Arabidopsis seeds were fixed to the seedbed with gum arabic were launched to the International Space Station in the space shuttle STS-128 (17A) on August 28, 2009. Plant Experiment Unit were installed in the Cell Biology Experiment Facility with desiccant boxes, and then the Space Seed experiment was started in the Japanese Experiment Module, named Kibo, which was part of the International Space Station, on September 10, 2009 by watering the seedbed and terminated 2 months later on November 11, 2009. On April 19, 2010, the Arabidopsis plants harvested in Kibo were retrieved and brought back to Earth by the space shuttle mission STS-131 (19A). The present paper describes the Space Seed experiment with particular reference to the development of the Plant Experiment Unit and its actual performance in Kibo onboard the International Space Station. Downlinked images from Kibo showed that the seeds had started germinating 3 days after the initial watering. The plants continued growing, producing rosette leaves, inflorescence stems, flowers, and fruits in the Plant Experiment Unit. In addition, the senescence of rosette leaves was found to be delayed in microgravity.     

A ground-based study for a shuttle BRIC experiment on gravity effects on gene expression.

A BRIC (Biological Research In a Canister) experiment to investigate the effects of reduced gravity at the molecular level using Arabidopsis has been initiated. In preparation for a space flight experiment, a series of ground-based studies were conducted. Results from these studies indicate that: 1) up to 20,000 seeds can be germinated on a 100 mm diameter Petri plate, 2) nylon membrane is the best surface for recovery of plant material after freezing, 3) depending on the age of the seedlings at the time of freezing, 20 to 40 g of tissue can be obtained from Petri plates that fit in a single canister; 4) tissue from one canister yields adequate amounts of RNA to perform differential display to isolate gravity-regulated genes. Our results indicate that the proposed BRIC experiment is feasible and can provide valuable information on the possible effects of microgravity on gene regulation.     

Simulated microgravity inhibits the proliferation of K562 erythroleukemia cells but does not result in apoptosis

Astronauts and experimental animals in space develop the anemia of space flight, but the underlying mechanisms are still unclear. In this study, the impact of simulated microgravity on proliferation, cell death, cell cycle progress and cytoskeleton of erythroid progenitor-like K562 leukemia cells was observed. K562 cells were cultured in NASA Rotary Cell Culture System (RCCS) that was used to simulate microgravity (at 15 rpm). After culture for 24 h, 48 h, 72 h, and 96 h, the cell densities cultured in RCCS were only 55.5%, 54.3%, 67.2% and 66.4% of the flask-cultured control cells, respectively. The percentages of trypan blue-stained dead cells and the percentages of apoptotic cells demonstrated no difference between RCCS-cultured cells and flask-cultured cells at every time points (from 12 h to 96 h). Compared with flask-cultured cells, RCCS culture induced an accumulation of cell number at S phase concomitant with a decrease at G0/G1 and G2/M phases at 12 h. But 12 h later (from 24 h to 60 h), the distribution of cell cycle phases in RCCS-cultured cells became no difference compared to flask-cultured cells. Consistent with the changes of cell cycle distribution, the levels of intercellular cyclins in RCCS-cultured cells changed at 12 h, including a decrease in cyclin A, and the increasing in cyclin B, D1 and E, and then (from 24 h to 36 h) began to restore to control levels. After RCCS culture for 12–36 h, the microfilaments showed uneven and clustered distribution, and the microtubules were highly disorganized. These results indicated that RCCS-simulated microgravity could induce a transient inhibition of proliferation, but not result in apoptosis, which could involve in the development of space flight anemia. K562 cells could be a useful model to research the effects of microgravity on differentiation and proliferation of hematopoietic cells.     

Preparation of the Biochip experiment on the EXPOSE-R2 mission outside the International Space Station

Biochips might be suited for planetary exploration. Indeed, they present great potential for the search for biomarkers – molecules that are the sign of past or present life in space – thanks to their size (miniaturized devices) and sensitivity. Their detection principle is based on the recognition of a target molecule by affinity receptors fixed on a solid surface. Consequently, one of the main concerns when developing such a system is the behavior of the biological receptors in a space environment. In this paper, we describe the preparation of an experiment planned to be part of the EXPOSE-R2 mission, which will be conducted on the EXPOSE-R facility, outside the International Space Station (ISS), in order to study the resistance of biochip models to space constraints (especially cosmic radiation and thermal cycling). This experiment overcomes the limits of ground tests which do not reproduce exactly the space parameters. Indeed, contrary to ground experiments where constraints are applied individually and in a limited time, the biochip models on the ISS will be exposed to cumulated constraints during several months. Finally, this ISS experiment is a necessary step towards planetary exploration as it will help assessing whether a biochip can be used for future exploration missions.     

The SOS-LUX-LAC-FLUORO-Toxicity-test on the International Space Station (ISS).

In the 21st century, an increasing number of astronauts will visit the International Space Station (ISS) for prolonged times. Therefore it is of utmost importance to provide necessary basic knowledge concerning risks to their health and their ability to work on the station and during extravehicular activities (EVA) in free space. It is the aim of one experiment of the German project TRIPLE-LUX (to be flown on the ISS) to provide an estimation of health risk resulting from exposure of the astronauts to the radiation in space inside the station as well as during extravehicular activities on one hand, and of exposure of astronauts to unavoidable or as yet unknown ISS-environmental genotoxic substances on the other. The project will (i) provide increased knowledge of the biological action of space radiation and enzymatic repair of DNA damage, (ii) uncover cellular mechanisms of synergistic interaction of microgravity and space radiation and (iii) examine the space craft milieu with highly specific biosensors. For these investigations, the bacterial biosensor SOS-LUX-LAC-FLUORO-Toxicity-test will be used, combining the SOS-LUX-Test invented at DLR Germany (Patent) with the commercially available LAC-FLUORO-Test. The SOS-LUX-Test comprises genetically modified bacteria transformed with the pBR322-derived plasmid pPLS-1. This plasmid carries the promoterless lux operon of Photobacterium leiognathi as a reporter element under control of the DNA-damage dependent SOS promoter of ColD as sensor element. This system reacts to radiation and other agents that induce DNA damages with a dose dependent measurable emission of bioluminescence of the transformed bacteria. The analogous LAC-FLUORO-Test has been developed for the detection of cellular responses to cytotoxins. It is based on the constitutive expression of green fluorescent protein (GFP) mediated by the bacterial protein expression vector pGFPuv (Clontech, Palo Alto, USA). In response to cytotoxic agents, this system reacts with a dose-dependent reduction of GFP-fluorescence. Currently, a fully automated miniaturized hardware system for the bacterial set up, which includes measurements of luminescence and fluorescence or absorption and the image analysis based evaluation is under development. During the first mission of the SOS-LUX-LAC-FLUORO-Toxicity-Test on the ISS, a standardized, DNA-damaging radiation source still to be determined will be used as a genotoxic inducer. A panel of recombinant Salmonella typhimurium strains carrying either the SOS-LUX plasmid or the fluorescence-mediating lac-GFPuv plasmid will be used to determine in parallel on one microplate the genotoxic and the cytotoxic action of the applied radiation in combination with microgravity. Either in addition to or in place of the fluorometric measurements of the cytotoxic agents, photometric measurements will simultaneously monitor cell growth, giving additional data on survival of the cells. The obtained data will be available on line during the TRIPLE-LUX mission time. Though it is the main goal during the TRIPLE-LUX mission to measure the radiation effect in microgravity, the SOS-LUX-LAC-FLUORO-Toxicity-test in principle is also applicable as a biomonitor for the detection and measurement of genotoxic substances in air or in the (recycled) water system on the ISS or on earth in general.     

3D information from 2D images recorded in the European Modular Cultivation System on the ISS

The European Modular Cultivation System (EMCS) on the ISS allows long-term biological experiments, e.g. on plants. Video cameras provide near real-time 2D images from these experiments. A method to obtain 3D coordinates and stereoscopic images from these 2D images has been developed and is described in this paper. The procedure was developed to enhance the data output of the MULTIGEN-1 experiment in 2007. One of the main objectives of the experiment was to study growth movements of the Arabidopsis plants and the effect of gravity on these. 3D data were important during parts of the experiment and the paper presents the method developed to acquire 3D data, the accuracy of the data, limitations to the technique and ways to improve the accuracy. Sequences of 3D data obtained from the MULTIGEN-1 experiment are used to illustrate the potential of this newfound capability of the EMCS. In the experiment setup, a positional depth accuracy of about ±0.4 mm for relative object distances and an absolute depth accuracy of about ±1.4 mm for time dependent phenomena was reached. The ability to both view biological specimens in 3D as well as obtaining quantitative 3D data added greatly to the scientific output of the MULTIGEN-1 experiment. The uses of the technique to other researchers and their experiments are discussed.     

Analysis of the space radiation doses obtained simultaneously at two different locations outside the ISS

Space weather and related ionizing radiation has been recognized as one of the main health concerns for the International Space Station (ISS) crew. The estimation of the radiation effect on humans outside the ISS requires at first order accurate knowledge of their accumulated absorbed dose rates, which depend on the global space radiation distribution, solar cycle and local variations generated by the 3D mass distribution surrounding the ISS. The R3DE (Radiation Risks Radiometer-Dosimeter for the EXPOSE-E platform) on the European Technological Exposure Facility (EuTEF) worked successfully outside of the European Columbus module between February 2008 and September 2009. A very similar instrument named R3DR for the EXPOSE-R platform worked outside the Russian Zvezda module of the ISS between March 2009 and August 2010. Both are Liulin-type detectors, Bulgarian-built miniature spectrometer-dosimeters. The acquired approximately 5 million deposited energy spectra from which the flux and absorbed dose rate were calculated with 10 s resolution behind less than 0.41 g cm⁻² shielding. This paper analyses the spectra collected in 2009 by the R3DE/R instruments and the long-term variations in the different radiation environments of Galactic Cosmic Rays (GCR), inner radiation belt trapped protons in the region of the South Atlantic Anomaly (SAA) and relativistic electrons from the Outer Radiation Belt (ORB). The R3DE instrument, heavily shielded by the surrounding structures, measured smaller primary fluxes and dose rates from energetic protons from the SAA and relativistic electrons from the ORB but higher values from GCRs because of the contribution from secondary particles. The main conclusion from this investigation is that the dose rates from different radiation sources around the International Space Station (ISS) have a large special and temporal dynamic range. The collected data can be interpreted as possible doses obtained by the cosmonauts and astronauts during Extra Vehicular Activities (EVA) because the R3DE/R instruments shielding is very similar to the Russian and American space suits average shielding (,  and ). Fast, active measurements are required to assess accurately the dose accumulated by astronauts during EVA.     

Changes in operational procedures to improve spaceflight experiments in plant biology in the European Modular Cultivation System

The microgravity environment aboard orbiting spacecraft has provided a unique laboratory to explore topics in basic plant biology as well as applied research on the use of plants in bioregenerative life support systems. Our group has utilized the European Modular Cultivation System (EMCS) aboard the International Space Station (ISS) to study plant growth, development, tropisms, and gene expression in a series of spaceflight experiments. The most current project performed on the ISS was termed Seedling Growth-1 (SG-1) which builds on the previous TROPI (for tropisms) experiments performed in 2006 and 2010. Major technical and operational changes in SG-1 (launched in March 2013) compared to the TROPI experiments include: (1) improvements in lighting conditions within the EMCS to optimize the environment for phototropism studies, (2) the use of infrared illumination to provide high-quality images of the seedlings, (3) modifications in procedures used in flight to improve the focus and overall quality of the images, and (4) changes in the atmospheric conditions in the EMCS incubator. In SG-1, a novel red-light-based phototropism in roots and hypocotyls of seedlings that was noted in TROPI was confirmed and now can be more precisely characterized based on the improvements in procedures. The lessons learned from sequential experiments in the TROPI hardware provide insights to other researchers developing space experiments in plant biology.     

Developing a vitamin greenhouse for the life support system of the International Space Station and for future interplanetary missions.

In order to evaluate the effects of gravity on growing plants, we conducted ground based long-term experiments with dwarf wheat, cultivar Apogee and Chinese cabbage, cultivar Khibinskaya. The test crops had been grown in overhead position with HPS lamp below root module so gravity and light intensity gradients had been in opposite direction. Plants of the control crop grew in normal position under the same lamp. Both crops were grown on porous metallic membranes with stable -1 kPa matric potential on their surface. Results from these and other studies allowed us to examine the differences in growth and development of the plants as well as the root systems in relation to the value of the gravity force influence. Dry weight of the roots from test group was decreased in 2.5 times for wheat and in 6 times - at the Chinese cabbage, but shoot dry biomass was practically same for both test and control versions. A harvest index of the test plants increased substantially. The data shows, that development of the plants was essentially changed in microgravity. The experiments in the space greenhouse Svet aboard the Mir space station proved that it is possible to compensate the effects of weightlessness on higher plants by manipulating gradients of environmental parameters (i.e. photon flux, matric potential in the root zone, etc.). However, the average productivity of Svet concerning salad crops even in ground studies did not provide more than 14 g fresh biomass per day. This does not provide a sufficient level of supplemental nutrients to the crew of the ISS. A cylindrical design of a space plant growth chamber (SPGC) allows for maximal productivity in presence of very tight energy and volume limitations onboard the ISS and provides a number of operational advantages. Productivity from this type of SPGF with a 0.5 kW energy utilization when salad growing would provide approximately 100 g of edible biomass per day, which would almost satisfy requirements for a crew of two in vitamin C and carotene and partly vitamin B group as well as rough fiber.     

Advances of Microgravity Sciences

Advances of microgravity sciences in China are introduced. The research works include ground-based study and space experiments. In the recent years, the main means still are theoretical analysis, numerical simulation, ground-based experiment, and short-time microgravity experiments of drop tower. Besides, many space experiment projects are arranged. SJ-10 recoverable satellite will carry out 19 scientific experiment projects. Nine of them are for microgravity Sciences. The other ways for space microgravity experiment are with the help of Chinese Shenzhou spacecraft, Chinese Tiangong space laboratory, and Chinese space station in the near future. The Chinese space station will become main platform of Chinese microgravity sciences experiment in space.      

The pleurodele, an animal model for space biology studies.

Pleurodeles waltl, an Urodele amphibian is proposed as a model for space biology studies. Our laboratory is developing three types of experiments in space using this animal: 1) in vivo fertilization and development ("FERTILE" project); 2) influence of microgravity and space radiation on the organization and preservation of specialized structures in the neurons and muscle cells (in vitro; "CELIMENE" PROJECT); 3) influence of microgravity on tissue regeneration (muscle, bone, epidermis and spinal cord).     

天宫二号碲化锌晶体生长

在天宫二号飞船综合材料实验炉六工位采用碲熔剂法生长了碲化锌晶体,生长时最高温度为800℃,以0.5mm·h-1的提拉速度向炉膛内部提拉生长晶体.飞行实验后,用相同实验参数在地面进行了对比实验.结果发现,空间样品尾部有一个非常大的橙色结晶区域（约10mm×6mm×2mm）,而地面生长样品中碲化锌晶体尺寸仅为约3mm×3mm×1mm,空间生长的碲化锌晶粒尺寸明显优于地面.空间微重力环境下,由于毛细作用,空间样品的塞子处有Te和ZnTe的外延膜生成.而地面生长的锭条在塞子处只有零星点状气相生产物.因此微重力条件有利于碲化锌晶体材料的生长.